Article ; Online: Immunomagnetic molecular probe with UHPLC-MS/MS: a promising way for reliable bronchial asthma diagnostics based on quantification of cysteinyl leukotrienes.
Journal of pharmaceutical and biomedical analysis
2013 Volume 81-82, Page(s) 108–117
Abstract: A sensitive and precise method for simultaneous quantification of cysteinyl leukotrienes (=cys LTs) - leukotriene C4 (=LTC4), leukotriene D4 (=LTD4) and leukotriene E4 (=LTE4) - essential biomarkers of bronchial asthma present in exhaled breath ... ...
| Abstract | A sensitive and precise method for simultaneous quantification of cysteinyl leukotrienes (=cys LTs) - leukotriene C4 (=LTC4), leukotriene D4 (=LTD4) and leukotriene E4 (=LTE4) - essential biomarkers of bronchial asthma present in exhaled breath condensate (=EBC) was developed. An immunomagnetic molecular probe was prepared by anchoring cysteinyl leukotrienes antibody on the surface of functionalized monodispersed magnetic particles and used to selectively isolate cys LTs from biological matrices - EBC, plasma and urine. Immobilization and the immunoaffinity capture procedures were optimized to maximize the amount of separated cys LTs, which were detected "off-beads" after acidic elution by UHPLC-ESI-MS/MS operated in a multiple reaction monitoring mode. The developed method was characterized with high precision ≤13.6% (intra-day precision determined as RSD) and ≤14.5% (inter-day precision determined as RSD), acceptable accuracy ≤18.5% (determined as RE), and high recovery of immunoseparation (≥93.1%) in aforementioned biological matrices. The applicability of the method was demonstrated on EBC, plasma and urine clinical samples of patients with various subtypes of bronchial asthma (occupational, steroid-resistant, moderate with and without corticosteroids therapy) and healthy subjects where reasonable differences in cys LTs concentration levels were found. Combining extremely selective immunomagnetic separation with highly sensitive and precise detection step, the developed method was used to aid diagnosis, predict the most effective therapy, and monitor the response to treatment. The detection of elevated inflammatory mediators (cys LTs) in EBC of subjects with relatively asymptomatic asthma and normal pulmonary function tests could offer a novel way for monitoring the lung inflammation and perhaps initiating treatment in an earlier stage. |
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| MeSH term(s) | Asthma/diagnosis ; Asthma/drug therapy ; Asthma/physiopathology ; Breath Tests ; Case-Control Studies ; Chromatography, High Pressure Liquid/methods ; Glucocorticoids/therapeutic use ; Humans ; Immunomagnetic Separation/methods ; Leukotriene C4/analysis ; Leukotriene D4/analysis ; Leukotriene E4/analysis ; Middle Aged ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization/methods ; Tandem Mass Spectrometry/methods |
| Chemical Substances | Glucocorticoids ; Leukotriene C4 (2CU6TT9V48) ; Leukotriene D4 (73836-78-9) ; Leukotriene E4 (75715-89-8) |
| Language | English |
| Publishing date | 2013-07 |
| Publishing country | England |
| Document type | Controlled Clinical Trial ; Journal Article ; Research Support, Non-U.S. Gov't ; Validation Studies |
| ZDB-ID | 604917-5 |
| ISSN | 1873-264X ; 0731-7085 |
| ISSN (online) | 1873-264X |
| ISSN | 0731-7085 |
| DOI | 10.1016/j.jpba.2013.03.026 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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